Soluble Human Cytomegalovirus gH/gL/pUL128–131 Pentameric Complex, but Not gH/gL, Inhibits Viral Entry to Epithelial Cells and Presents Dominant Native Neutralizing Epitopes

• DOI: 10.1074/jbc.m115.652230
• Publication Date: 2015-05-07T01:32:50Z
• Authors: John W. Loughney; Richard R. Rustandi; Dai Wang; Matthew C. Troutman; Lawrence W. Dick; Guanghua Li; Zhong Liu; Fengsheng Li; Daniel C. Freed; Colleen E. Price; Van M. Hoang; Timothy D. Culp; Pete A. DePhillips; Tong-Ming Fu; Sha Ha
• Keywords: 0301 basic medicine; Membrane Glycoproteins; Cytomegalovirus; Epithelial Cells; Virus Internalization; Antibodies, Viral; Antibodies, Neutralizing; 3. Good health; Epitopes; 03 medical and health sciences; Viral Envelope Proteins; Cricetinae; Cytomegalovirus Infections; Animals; Humans; Protein Binding

Congenital infection of human cytomegalovirus (HCMV) is one the leading causes nongenetic birth defects, and development a prophylactic vaccine against HCMV high priority for public health. The gH/gL/pUL128–131 pentameric complex mediates entry into endothelial epithelial cells, it major target neutralizing antibody responses. To better understand mechanism by which antibodies interact with epitopes resulting in viral neutralization, we expressed purified soluble gH/gL from Chinese hamster ovary cells to >95% purity. gH/gL, exists predominantly as (gH/gL)2 homodimer molecular mass 220 kDa solution, has stoichiometry 1:1 pI 6.0–6.5. 160 kDa, 1:1:1:1:1, 7.4–8.1. complex, but not adsorbs 76% activities hyperimmune globulin, consistent earlier reports that most potent blocking are unique complex. Functionally, blocks culture. Our results highlight importance design emphasize necessity monitor integrity during manufacturing process.